Effect of slash removal on Gremmeniella abietina incidence on Pinus sylvestris after clear-cutting in northern Sweden

The Gremmeniella abietina outbreak in Sweden in 2001-2003 forced forest owners to sanitary clear-cut large areas of middle-aged Pinus sylvestris stands. There is, however, little knowledge of effective reforestation of P. sylvestris on G. abietina-infected sites. Gremmeniella abietina disease incidence on P. sylvestris seedlings planted in 2003 was studied with and without (control) removal of infected P. sylvestris slash. Removed slash was piled in stacks around the regeneration plots. The seedlings were planted within 1 year after sanitation felling on three sites in northern Sweden. One year after planting, G. abietina pycnidia were found on 32% of the control seedlings and total infection, including stem cankers, reached 44%. Total and G. abietina-induced mortality was 15 and 10%, respectively. The method of removing and piling the infected slash reduced the number of infected seedlings by 50% and seedling mortality by 27% 1 year after planting, compared with control. Consequently, even if there is a clear sanitation effect of removing infected slash to the sides of the regeneration area, it does not eradicate the infection source from the stands. Postponed planting, slash burning or complete removal of the infected slash is needed to minimize the infection risk. The positive correlation found between slash coverage and infection rate indicates that clear-cuts with large amounts of infected slash should be given priority for slash treatment.     

Susceptibility of six pine species to Gremmeniella abietina isolates from Spain

Inoculation experiments were performed in order to evaluate the virulence of Gremmeniella abietina isolates from Spain on the main pine species planted in the Iberian Peninsula, as well as the influence of seedling age on this virulence. Two different experiments were carried out with four isolates of G. abietina from Spain. The greenhouse experiments consisted of seedling inoculations. One‐ and 2‐year‐old seedlings of the following five pine species were used: Pinus halepensis, Pinus pinea, Pinus pinaster, Pinus sylvestris and Pinus uncinata; also, 1‐year‐old seedlings of P. nigra were inoculated. The relative necrosis length (RNL) caused by the pathogen after 130 days was used as a response variable. The laboratory experiments were performed on 2‐ to 6‐year‐old internodes of the above pine species excluding P. uncinata. The necrosis length after 6 weeks of incubation was measured. The results have shown that all G. abietina isolates were pathogens on seedlings of these six pine species and seedlings of P. halepensis were consistently the most susceptible ones, although it is important to take into account that all the isolates used in the present work were isolated from P. halepensis, the only pine species in Spain where G. abietina has been recovered up to now. The susceptibility of the other pine species depended on the age of the seedlings.     

Resistance of Pinus contorta to the European race of Gremmeniella abietina

Pinus contorta seedlings, together with Pinus resinosa and Pinus banksiana seedlings, were planted adjacent to 25‐year‐old red pine trees infected by the European (EU) race of Gremmeniella abietina. Resistance to this race was assessed over 5 years. All P. resinosa seedlings were dead after that period while 65% of P. contorta and 86% of P. banksiana seedlings appeared resistant to the disease. The tip blight that occurred on P. contorta was slightly longer than that observed on P. banksiana. In microscopy, one, two, or even more suberized boundaries were seen to be initiated near the surface of the shoot at the base of healthy needles where they extended downward in the direction of the vascular cambium. Suberized boundaries occasionally crossed the xylem and joined together in the pith region to form continuous barriers around necrotic tissues. However, in most cases, these suberized barriers were not continuous across the shoot and compartmentalization was then completed by other barriers mainly constituted of parenchyma cells and xylem tracheids that accumulated phenolic compounds. Meristematic‐like cells were observed adjacent to the necrophylactic periderm. Tissue regeneration, restoration of cambial activities and formation of traumatic resin canals also seemed to be associated with the defence system of P. contorta against the EU race of G. abietina.     

Susceptibility of different provenances of Pinus sylvestris,Pinus Contorta and Picea abies to Gremmeniella abietina

In a randomized block factorial experiment, 1200 seedlings from four provenances, each of Pinus sylvestris, Pinus contorta and Picea abies were inoculated with conidia (2 × 10⁴ and 1 × 10⁶ conidia/seedling) of Gremmeniella abietina (Brunchorstia pinea), isolated from P. contorta plantations in northern Sweden. A further 600 seedlings were left as controls. The occurrence of symptoms and the extension of dead tissues on the annual shoots were recorded 13 months after inoculation. Only the higher spore dose resulted in significant infection. P. sylvestris and P. contorta seedlings were equally susceptible (53% infected), and significantly more infected than P. abies seedlings (39%). The annual shoots of P. sylvestris and P. abies were affected to 43% and 37% of their length, which was significantly more than 15% of the length of P. Contorta shoots. The frequency of affected seedlings differed between the most southern and northern provenances of P. sylvestris and P. abies, and between the northwestern and the south-eastern provenance of P. contorta Twenty-six months after inoculation, a higher proportion of P. contorta seedlings than P. sylvestris and P. abies seedlings had recovered, and a lower proportion of P. contorta than P. sylvestris and P. abies seedlings had died. This paper discusses why P. sylvestris was found to be more susceptible to G. abietina than P. contorta in this experiment, while the reverse is found in plantations in northern Sweden.     

Effects of associated fungi Sclerophoma pythiophila and Cenangium ferruginosum on Gremmeniella abietina dieback in Spain

The interactions between Gremmeniella abietina and either Sclerophoma pythiophila or Cenangium ferruginosum, fungi frequently isolated from diseased twigs along with G. abietina, were studied under laboratory (dual cultures) and greenhouse conditions (double‐inoculations). Virulence of each species was also evaluated in greenhouse experiments by means of single‐inoculations. In vitro interactions were assessed on Petri dishes containing malt agar with pine needle extract, and greenhouse experiments were performed on 1‐year‐old Pinus halepensis seedlings. In vitro growth of G. abietina was inhibited by both fungi when grown in dual culture. In single‐inoculations, G. abietina caused the greatest necrosis length on P. halepensis seedlings, followed by S. pythiophila, whereas C. ferruginosum did not cause significant necrosis. In double‐inoculations, C. ferruginosum was able to reduce the length of necrosis caused by G. abietina on the P. halepensis seedlings. In contrast, necrosis length was greater in seedlings inoculated with both S. pythiophila and G. abietina than in those inoculated with G. abietina alone. Therefore, S. pythiophila seems to play a role in disease expression caused by G. abietina on P. halepensis in Spain.     

Infection experiments with Gremmeniella abietina on seedlings of Norway spruce and Scots pine

Conidia of Gremmeniella abietina infected and caused disease symptoms in annual shoots of both Scots pine (Pinus sylvestris) and Norway spruce (Picea abies) seedlings. In Norway spruce shoots the infection remained largely latent, with only a few seedlings showing symptoms. Mycelial growth inside the shoots was faster in Scots pine than in Norway spruce and was favoured by low temperature in both hosts. The shoots of Norway spruce seedlings had higher endophyte populations than those of Scots pine, and the populations were decreased by low temperatures. Reductions in the normal epiphytic or endophytic flora by acid mist treatments seemed to favour the development of G. abietina.     

Inoculation of young pine seedlings with Peridermium pini from north-east Scotland

Seedlings of Pinus sylvestris, P. nigra ssp. laricio, and P. mugo vars mughus, rostrata and pumilio were inoculated at their cotyledon stage with aeciospores of Peridermium pini from north-east Scotland. Infection was determined by the presence of both mycelia and haustoria in free-hand or paraffin sections. In the second growing season, infected seedlings showed symptoms such as stem-swelling, development of spermogonia and aecia, and death. In a second experiment, young P. sylvestris seedlings from seven seed sources collected in Great Britain were inoculated and the infection were examined after 6 weeks. In the samples of 50 seedlings from each source, 30–70% seedlings were infected.     

Comparison of Gremmeniella abietina isolates from Pinus sylvestris and Pinus contorta in terms of conidial morphology and host colonization

Gremmeniella abietina isolates from Pinus contorta in northern Sweden produced, in vitro, shorter conidia with fewer septa compared with isolates from Pinus sylvestris in the southern part of the country. After mycelial inoculation of shoots with G. abietina isolates from both host species, the resulting necroses were longer in P. sylvestris than in P. contorta. Keeping seedlings in artificial mild winter climate or detaching shoots from the seedling before inoculation caused longer necroses. No host specificity in colonization was found. Isolates from P. sylvestris caused longer necroses than did isolates from P. contorta, and both types of isolates caused longer necroses in P. sylvestris than in P. contorta. The differences found between the two G. abietina populations probably reflect regional variation in the fungus.     

Virulence of two Finnish Gremmeniella abietina types (A and B)

Thirteen Finnish Gremmeniella abietina isolates of types A and B were compared for differences in their virulence. Three kinds of inoculations were made: one with conidia on young Scots pine (Pinus sylvestris) seedlings and two with mycelia on the stems and shoots of young Scots pine trees. Inoculations with conidia were carried out in August 1992 and inoculations with mycelia were carried out six times between 11 August and 20 October, 1992. The experiments were evaluated in the late spring and early summer of 1993. The results showed that there was a difference in virulence between the two types. In the conidial inoculations type A infected 34.7% and type B infected 11.0% of the inoculated seedlings. For mycelial inoculations with type A the mean canker (stems) and necrosis (shoots) lengths were 19.3 mm and 8.6 mm longer, respectively, than with type B inoculations. In shoot inoculations there also was a clear difference between the two types in the number of such inoculations where no symptoms were observed. For type B shoot inoculations there was no fungal growth in 21.5% whereas for type A inoculations the figure was only 3.7%.     

Natural regeneration on planted clear‐cuts in boreal Sweden

Natural regeneration was registered on 97 clear‐felled and scarified sites in northeastern Sweden ten growing seasons after planting. On each site, three fix plots were laid out at planting and the planted seedlings were mapped. Ten years later all healthy non‐planted seedlings were registered in an anulus of size 15.7 m². Non‐planted seedlings were also observed in four 4 m² squares on each fix‐plot to estimate the zero‐square frequency. Species were classified as Pinus sylvestris, L., Picea abies (L) Karst. and deciduous species (Betula pendulajpubescens Roth./Ehrh., Populus tremula L., Alnus incana (L) Moench.). On average, 7 133 non‐planted seedlings ha^‐1 were found, of which 29% were conifers. Among the conifers, 71% were P. abies. More than 2 500 conifer non‐planted seedlings ha^‐1 were found on 27% of the plots. The number of non‐planted seedlings was correlated to the presence of neighbouring seed‐producing stands within 100 m and decreased with rising altitude. Pinus sylvestris seedlings were more frequent on dry soils while the deciduous seedlings were more frequent on wet sites. In the 4 m² squares, the zero‐square frequency was 29% if both conifers were and deciduous seedlings were considered and 63% if only conifers were considered. Mean height of the tallest seedling per species and square was 62 cm for P. sylvestris, 42 cm for P. abies and 107 cm for the deciduous seedlings. Key words: Alnus incana, Betula, broad‐leaves, deciduous, Norway spruce, Picea abies, Pinus sylvestris, Populus tremula, reforestation, Scots pine.     

Tree mortality caused by Gremmeniella abietina in a subalpine afforestation in the central Alps and its relationship with duration of snow cover

Tree survival and causes of mortality were studied in an experimental afforestation in the upper subalpine forest zone in the Swiss Alps. A total of 59.8% of Pinus cembra and 45.6% of Pinus mugo were killed by Gremmeniella abietina during the first 20 years after planting, compared to 1.5% of Larix deddua trees. The mortality rates caused by G. abietina were highly correlated with the duration of snow cover in spring. Tree losses were lowest at sites where the snow melted early and highest at sites where the snow ablation was delayed in spring. Tree mortality varied greatly between years. In the year after the coldest summer of the observation period mortality due to G. abietina infections was highest, suggesting high susceptibility of trees in poor condition. Phaddium infestans, which was the second most important factor for mortality in P. cembra, killed trees irrespective of their condition. Other biotic and abiotic causes of tree mortality had negligible influence compared with the impact of G abietina and P. infestans. Excluding anthropogenic impacts, the diverse spatial pattern of forested and treeless sites close to the subalpine timberline may predominantly result from the action of parasitic fungi, depending on the ablation pattern of the snow cover in spring.     

Effects of sulphuric acid and heavy-metal deposition on frequency of asymptomatic infections of Gremmeniella abietina in Scots-pine seedlings

The isolation frequency of Gremmeniella abietina was measured from asymptomatic 4-year-old Scots-pine seedlings. The seedlings had been subjected to simulated sulphuric deposition (pH 3.1 or 4.1) or control treatments (irrigation with spring water (pH 6), or no irrigation) during the three previous growing seasons. The seedlings were planted in soil originating from two localities with different backround levels of copper (Cu) and nickel (Ni), and the soil was further amended with Cu (63.3 mg/1 of soil) and Ni (20.5 mg/1 of soil), or a combination of the two. The seedlings were inoculated with conidia of G. abietina during their third growing season in the field. Asymptomatic infections of G. abietina were recovered frequently from seedlings that had received water irrigation. The addition of sulphuric acid to the irrigation water, and/or the lowered pH (pH 3.1 and 4.1) resulting from this addition, mitigated the effect of water application. The effect of Ni-treatment on the recovery of G. abietina depended on the irrigation treatment; with no irrigation, Ni-treatment increased isolation frequency, but, in irrigated seedlings the response was the opposite. Only 10.6% of the seedlings had symptoms caused by G. abietina; most of the seedlings had become infected before the application of the various treatments.     

RAPD variation in Gremmeniella abietina attacking Pinus sylvestris and Pinus contorta in northern Sweden

Genomic DNA from 81 isolates of Gremmeniella abietina collected from eleven plantations each of Pinus sylvestris and Pinus contorta in northern Sweden was studied using RAPD markers. The DNA variation between and within populations and the race and type distribution of G. abietina populations, causing symptoms similar to those of the North American race, were studied. The degree of genetic similarity was greater among G. abietina isolates from the same geographical areas than among isolates from different geographical areas, regardless of whether they were isolated from P. sylvestris or P. contorta. RAPD variation was greatest in the central parts of northern Sweden, suggesting that sexual reproduction has been somewhat more important there than further north or south. Only the RAPD fragments characteristic of the EU race of G. abietina were found in the material tested. The RAPD pattern described as characteristic of the northern type within the EU race was identified in 62% of the isolates. Divergence from the expected profile was due to differences in occurrence of fragments OPA12-1400 and 12-1500. This indicates that this part of the RAPD profile cannot be treated as diagnostic for the northern type. A conclusion of practical importance is that there is a considerable risk of G. abietina spreading from infected P. contorta plantations to adjacent areas with indigenous P. sylvestris regeneration, and vice versa, owing to the indicated lack of host-specificity of the pathogen. It is possible, however, that host-specific strains exist, but do not differ in their RAPD profiles.     

From the Editor

Abstract

In 2001 large areas of Pinus sylvestris L. stands in Sweden were attacked by Gremmeniella abietina (Lagerb.) Morelet. The resultant damage was assessed in five genetic field trials and one clonal seed orchard, with the aims of quantifying the genetic variation in tree responses to G. abietina, studying the genetic relationship between G. abietina susceptibility and growth traits, and examining the implications of the results for P. sylvestris breeding. A comparison of G. abietina damage between plus-tree progenies and seed stand check-lots in each of four tree classes (dominant, co-dominant, subordinated and suppressed) found no significant differences except in the suppressed tree class, where plus-tree progenies were damaged less severely than trees from the seed stands. Narrow- and broad-sense heritability for G. abietina damage traits were in the range 0.22–0.42. Genetic correlations between G. abietina damage and growth traits were weak and not significantly different from zero. The results imply that selection for growth capacity in P. sylvestris breeding programmes using plus-tree selection or field progeny tests does not increase susceptibility to G. abietina. In addition, there is scope for selecting genotypes with increased resistance if this is required in the future.     

Development of fruiting bodies of large tree type of Gremmeniella abietina var. abietina and timing of infection on Scots pine inn northern Finland

The formation and maturing of the large tree type Gremmeniella abietina var. abietina fruiting bodies and their sporulation were investigated for 3 years on Scots pine (Pinus sylvestris) in northern Finland. This was done by monthly assessment of shoots in the field and in the laboratory. Infection caused by G. abietina var. abietina was dated on Scots pine by monthly covering with pollination bags and exposing branches during the growing season. Pycnidia appeared between August and September, 1 year after infection, and they started to release conidia between late June and early July, 2 years after infection. Fresh pycnidia and microconidia were formed during the following August and September in the infected shoots. The causal large tree type of G. abietina var. abietina did not produce apothecia on branches within 3 years of infection. Monthly covering and exposing branches showed that infection took place mainly between June and July.     

Pathogenicity of Heterobasidion annosum (Fr.) Bref. sensu stricto on coniferous tree species in Turkey

Two‐year‐old seedlings of Pinus brutia, P. brutia var. eldarica, Pinus pinea and 3‐year‐old seedlings of Pinus radiata, Pinus sylvestris, Pinus nigra and Cedrus libani were inoculated on the lower stem with isolates of Heterobasidion annosum s.s. collected from the Black Sea and Mediterranean regions of Turkey. In total, 315 seedlings were inoculated in April 2014 and incubated in a growth chamber for 7 weeks at 18–20°C. All isolates were pathogenic on the seven different hosts and had the ability to grow in living sapwood. The isolates had a greater growth on C. libani, P. sylvestris and P. radiata seedlings compared to plants of the other species tested. The least affected species were P. brutia and P. nigra. The isolates originating from the Black Sea region caused longer lesions on the hosts. Overall mortality during 7 weeks of incubation was 4%.     

Survival and vitality of Gremmeniella abietina on Pinus sylvestris slash in northern Sweden

Survival and vitality of Gremmeniella abietina on Pinus sylvestris slash was studied in northern Sweden during 2003 and 2004. Once a month between September 2003 and April 2004, two to three trees were cut down and debranched. Shoots with pycnidia were sampled at the felling date and then at every consecutive month. The percentage of germinated conidia from each shoot was calculated after 24, 48 and 72 h incubation. The vitality of G. abietina pycnidia in the slash remained high the whole period. Intact pycnidia were found on slash several months after the time of conidial sporulation, which indicates that new pycnidia may be produced on dead pine branches. Sampling of shoots from slash on 13‐ to 18‐month‐old clear‐cuts showed conidial germination capacity as high as in pycnidia collected in fresh slash. Due to survival of G. abietina in slash it is recommended to postpone planting of P. sylvestris seedlings in northern boreal areas to the third vegetation period after sanitary clear‐cuts.     

Enzymatic interactions between Gremmeniella abietina var. abietina,European race,and two resistant hosts,Pinus banksiana and P. contorta

Ultrastructural observations showed that the European race of Gremmeniella abietina var. abietina can secrete several cell wall–degrading enzymes that help the invasion of Pinus banksiana and P. contorta shoot tip tissues. Alterations in the content or distribution of cellulose were obvious, indicating that colonization might be achieved primarily through the action of glucanases. Polygalacturonases seemed more abundant in the cortex and the phloem of the stem, suggesting they are among the first enzymes secreted by the pathogen during infection. Production of laccases and peroxidases was also revealed but always close to fungal cells or in areas where host wall degradation was obvious. In places where there was an accumulation of phenolic compounds in the infected zones of the shoot, particularly in or near the transition zone, immunolocalization of both enzymes was limited or absent. Laccases and polygalacturonases were regularly detected over the extracellular sheath of G. abietina, and this indicates that the sheath could play a significant role during host wall degradation. Finally, examination of G. abietina wall constituents, such as chitin and β‐1,3‐glucans, suggests that the composition of the pathogen wall changes during the infection process. This change may be associated with the secretion of fungal enzymes but could also be linked to host reactions altering the integrity of pathogen cell walls.     

Importance of Site Type and Tree Species on Disease Incidence of Gremmeniella abietina in Areas with a Harsh Climate in Northern Sweden

The importance of site type and tree species on disease incidence of Gremmeniella abietina was investigated. Blocks of four conventional regenerations of Pinus contorta and Pinus sylvestris, situated on sites formerly supporting pure stands of P. sylvestris and Picea abies respectively, were examined for the occurrence of G. abietina. Trees planted on spruce sites were found to be more severely diseased by G. abietina than trees planted on pine sites. Pinus contorta was more severely attacked by G. abietina than P. sylvestris on both site types. The results show that plantations of P. contorta and P. sylvestris in areas originally covered with pure spruce forests run a high risk of damage by G. abietina.     

Gremmeniella abietina,Phacidium infestans,and other causes of damage in alpine,young pine plantations in Norway

Field studies of damage and mortality were carried out through 16 years in six young plantations, mostly of Pinus contorta, but also P. sylvestris and P. sibirica. Differences between pine species, P. contorta provenances, sites, and soil treatments are reported and discussed, as are influences of environment, climatic conditions, snow cover, and nutrient deficiency. Gremmeniella abietina was by far the most important cause of mortality; there were great differences between the sites as to development and seriousness. Other important causes of mortality and damage were Phacidium infestans, elk (Alces alces L.), voles, and snow pressure.